Background Many patients with heart failure (HF) have cognitive deficits, including memory loss. Objectives To evaluate the efficacy of a cognitive training intervention on memory (primary outcome), working memory, psychomotor speed, executive function, and performance of cognitive activities and instrumental activities of daily living (IADLs). Methods Forty patients with HF were randomly assigned to the computerized plasticity-based cognitive training intervention called Brain Fitness and to the health education active control intervention. Advanced practice nurses made weekly home visits to assess symptoms and monitor intervention adherence. Patients completed demographic and clinical data (baseline), neuropsychological tests (baseline, 8, and 12 weeks), and measures of cognitive and IADLs performance (baseline and 12 weeks)and satisfaction (12 weeks). Results Linear mixed models analyses indicated a significant group by time interaction for delayed recall memory (p = .032) and a significant time effect for total (list learning) (p < .001) and delayed (p = .015) recall memory, psychomotor speed (p = .029), and performance of IADLs (p = .006). Intervention adherence and patient satisfaction were high. Conclusions To our knowledge, this was thefirst test of Brain Fitness in HF. Although it was a preliminary study with limitations, results support the need for a largerrandomized, controlled trialto determine whether the memory loss of HF is amenable to plasticity-based interventions.

The gene for a novel extracellular metalioprotease was cloned, and its nucleotide sequence was determined. The gene (mpr) encodes a primary product of 313 amino acids that has little similarity to other known Bacillus proteases. The amnmo acid sequence of the mature protease was preceded by a signal sequence of approximately 34 amino acids and a pro sequence of 58 amino acids. Four cysteine residues were found in the deduced amino acid sequence of the mature protein, indicating the possible presence of disulfide bonds. The mpr gene mapped in the cysA-arol region of the chromosome and was not required for growth or sporulation.The gram-positive, spore-forming bacterium Bacillus subtilis produces and secretes proteases, esterases, and other types of exoenzymes at the end of the exponential phase of growth (15). The principal extracellular proteolytic enzymes, alkaline (subtilisin) and neutral (metallo-) proteases, are encoded by the apr and npr genes, respectively (11,24,26,29). In addition, the genes for two minor extracellular proteases, epr (21) and bpr (encoding bacillopeptidase F; A. Sloma, G. A. Rufo, Jr., C. F. Rudolph, B. J. Sullivan, K. A. Theriault, and J. Pero, submitted for publication) have been identified. Strains bearing null mutations in these four protease genes and the major intracellular protease gene isp-i (12) still produce extracellular protease activity. The majority of the residual protease activity can be attributed to a novel cysteine-containing metalloprotease (Mpr) that has been purified to apparent homogeneity (17). Here we report the cloning of the gene encoding this cysteine-containing protease. MATERIALS AND METHODSBacterial strains and plasmids. B. subtilis strains are listed in Table 1. Plasmid pBs81/6 is a derivative of pBD64 (8). Plasmids pBR322 and pUC19 were used for cloning into Escherichia coli DH5 cells obtained from Bethesda Research Laboratories, Inc., Gaithersburg, Md. The cat gene was obtained from plasmid pMI1101 (30), and the ble gene was obtained from pUB110 (20). B. subtilis strains were grown on tryptose blood agar base or minimal glucose medium and were made competent by the procedure of Anagnostopoulos and Spizizen (1). Selection for phleomycin resistance was carried out on tryptose blood agar base by the overlayer method after a 1.5-h delay at 37°C to allow for expression.The final concentration of phleomycin was 2 jig/ml. Plasmid DNA from B. subtilis and E. coli was prepared by the alkaline lysis method of Birnboim and Doly (3). Plasmid DNA transformation in B. subtilis was performed as described by Gryczan et al. Amino acid sequence determination. The N-terminal amino acid sequence of purified Mpr was determined by automated sequential Edman degradation with subsequent identification and quantification of phenylthiohydantoin-labeled amino acids by reverse-phase high-performance liquid chromatography. Additional amino acid sequences were obtained from internal fragments of Mpr generated by trypsin digestion. The purified enzyme was incubated with trypsin, and t...

We have purified a minor extracellular serine protease from Bacillus subtilis. Characterization of this enzyme indicated that it was most likely the previously reported enzyme bacillopeptidase F. The amino-terminal sequence of the purified protein was determined, and a "guess-mer" oligonucleotide hybridization probe was constructed on the basis of that sequence. This probe was used to identify and clone the structural gene (bpr) for bacillopeptidase F. The deduced amino acid sequence for the mature protein (496 amino acids) was preceded by a putative signal sequence of 30 residues and a putative propeptide region of 164 amino acids. The bpr gene mapped near pyrD on the chromosome and was not required for growth or sporulation.

We have isolated and characterized two minor extracellular proteases from culture supernatants of a strain of Bacillus subtilis containing deletion mutations of the genes for the extracellular proteases subtilisin (apr) and neutral protease (npr) and a minor extracellular protease (epr) as well as intracellular serine protease-I (isp-1). Characterization studies have revealed that one of these enzymes is the previously described protease bacillopeptidase F. The second enzyme, the subject of this report, is a novel metalloprotease, which we designate Mpr. Mpr is a unique metalloprotease that has been purified to apparent homogeneity by using both conventional and high-performance liquid chromatography procedures. Mpr has a molecular mass of -28 kilodaltons on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a basic isoelectric point of 8.7. The enzyme showed maximal activity against azocoll at pH 7.5 and 50°C. Mpr was inhibited by dithiothreitol and a combination of j8-mercaptoethanol and EDTA. Activity was moderately inhibited by I-mercaptoethanol and EDTA alone as well as by cysteine and citrate and only marginally by phosphoramidon 1,10-phenanthroline and N-[N-(L-3-trans-carboxyoxiran-2-carbonyl)-L-leucyl]-agmatine. Mpr was not inhibited by phenylmethylsulfonyl fluoride. In addition, Mpr showed esterolytic but not collagenolytic activities. Our studies suggest that Mpr is a secreted metalloprotease containing cysteine residues that are required for maximal activity.Sporulation in Bacillus subtilis is a tightly regulated developmental process involving alterations in protein synthesis and intermediary metabolism (5,6,20). At the end of exponential growth and paralleling the onset of sporulation, there is increased production and secretion of extracellular proteases.Two prominent proteases produced during the onset of sporulation are the alkaline serine protease or subtilisin (12, 22) and neutral (metallo-) protease (23)(24)(25). The combined activities of these two enzymes account for 96 to 98% of the total protease activity present in culture supernatants of wild-type sporulating cells (8). Efforts to clone and delete the genes coding for subtilisin (8, 21) and neutral protease (28) have made possible the study of minor proteases (1,13,14) that contribute to the residual activity present in double mutants of B. subtilis incapable of producing either protease. For example, work in our laboratory has identified and resulted in the cloning of a minor extracellular protease gene (epr; 17) from a strain of B. subtilis deleted for the subtilisin (apr) and neutral protease (npr) genes.

This investigation was designed to examine self-esteem and depression in diabetic adolescent girls. One hundred nondiabetic girls age 12-16 and 105 diabetic girls age 12-16 were administered the Rosenberg Self-Esteem Scale and the Beck Depression Inventory. Results indicated no significant difference between diabetic and nondiabetic girls in self-esteem scores. Diabetic girls showed significantly more depression than nondiabetic girls. Close examination of results revealed that, in fact, diabetic and nondiabetic adolescent girls were very similar. A major finding was that depression in the diabetic group was expressed primarily through physiologic symptoms of depression as seen in the vital depression scores, rather than through the pessimism, indecision-inhibition, or self-debasement measures of depression. Results were interpreted to mean that diabetic girls did not manifest deeper depression than nondiabetic girls but, rather, a greater awareness of their physiologic status. Diabetes emerged as a focus for the expression of normal adolescent conflicts. The importance of integrating developmental issues into the treatment plans for diabetic patients is emphasized.